【摘要】：植物紫色酸性磷酸酶(purple acid phosphatase,PAP)在植物響應(yīng)低磷脅迫過程中起重要作用,為了驗證馬尾松(Pinus massoniana)紫色酸性磷酸酶(PmPAP1)基因在磷吸收上的功能,本研究通過農(nóng)桿菌(Agrobacterium tumefaciens)介導(dǎo)法轉(zhuǎn)化煙草(Nicotiana tabacum),獲得25個轉(zhuǎn)基因株系,RT-PCR及qRTPCR分析表明,PmPAP1在13個株系中高表達(dá);對其中2個高表達(dá)株系(Line-10和Line-13)進(jìn)行不同濃度磷處理,結(jié)果表明,缺磷可以誘導(dǎo)轉(zhuǎn)基因植株中PmPAP1基因在根和葉中上調(diào)表達(dá);低磷條件下,轉(zhuǎn)基因植株根系及葉片酸性磷酸酶(acid phosphatase,APase)活性分別為野生型(wild type,WT)的1.27和1.25倍;轉(zhuǎn)基因植株總磷與無機(jī)磷含量分別比WT提高了28.32%、45.48%(Line-10)與36.45%、62.31%(Line-13)。除此之外,缺磷條件下轉(zhuǎn)基因植株過氧化物酶(peroxidase,POD)、超氧化物歧化酶(superoxide dismutase,SOD)均較WT顯著提高(P0.05),而丙二醛(malondialdehyde,MDA)含量則顯著下降(P0.05)。磷脅迫對植株生長影響表明,低磷條件下轉(zhuǎn)基因植株總生物量、根系生物量及根冠比均顯著提高(P0.05)。而在正常和高磷條件下,轉(zhuǎn)基因與野生型煙草的酶活性、丙二醛含量、磷含量和生物量等與均無明顯差異。因此,高表達(dá)PmPAP1基因能顯著提高煙草植株在低有效磷下磷的利用能力,進(jìn)而增強(qiáng)耐低磷脅迫能力。本研究為深入了解馬尾松耐低磷的分子機(jī)制與創(chuàng)制耐低磷煙草新種質(zhì)提供了理論依據(jù)。
[Abstract]:Plant purple acid phosphatase (purple acid) plays an important role in response to low phosphorus stress. In order to verify the function of (Pinus massoniana) purple acid phosphatase (PmPAP1) gene in phosphorus uptake of Pinus massoniana. RT-PCR and qRTPCR analysis of 25 transgenic lines obtained by Agrobacterium tumefaciens (Agrobacterium tumefaciens) mediated transformation of tobacco (Nicotiana tabacum), showed that PmPAP1 was overexpressed in 13 lines, and 2 of them (Line-10 and Line-13) were treated with different concentrations of phosphorus. Phosphorus deficiency could induce the expression of PmPAP1 gene in the roots and leaves of transgenic plants, and the activity of acid phosphatase apase in roots and leaves of transgenic plants was 1.27 and 1.25 times higher than that of wild type (wild type WT) under the condition of low phosphorus. Compared with WT, the total phosphorus and inorganic phosphorus contents of transgenic plants were increased by 45.48% (Line-10) and 62.31% (Line-13), respectively. In addition, the content of peroxidase (POD) and superoxide dismutase (superoxide) in transgenic plants were significantly higher than that in WT (P0.05), while the content of malondialdehyde (malondialdehyde) decreased significantly (P0.05). The effects of phosphorus stress on plant growth showed that the total biomass root biomass and root-shoot ratio of transgenic plants were significantly increased under low phosphorus condition (P0.05). However, there was no significant difference in enzyme activity, malondialdehyde content, phosphorus content and biomass between transgenic tobacco and wild type tobacco under normal and high phosphorus conditions. Therefore, the high expression of PmPAP1 gene could significantly improve the phosphorus utilization ability of tobacco plants under low available phosphorus, and then enhance the ability of tolerance to low phosphorus stress. This study provided a theoretical basis for understanding the molecular mechanism of low phosphorus tolerance of Pinus massoniana and for creating new low phosphorus tolerant tobacco germplasm.
【作者單位】： 貴州大學(xué)農(nóng)業(yè)生物工程研究院/生命科學(xué)學(xué)院/山地植物資源保護(hù)與種質(zhì)創(chuàng)新教育部重點(diǎn)實驗室;貴州大學(xué)林學(xué)院;
【基金】：國家高技術(shù)研究發(fā)展計劃(863)項目(No.2011AA10020301) 貴州省重大專項(No.20126011-1)
【分類號】：S791.248

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